Abstract: Since 80’s,the isolation and biological studies of sperm cells in angiosperms have been an important research area in plant fertilization engineering. Based on the isolation of vialbe sperm of Liriodendron chinense, the mass isolation of viable sperm cells form the its pollen was reported. With sugar density gradient centrifugation method, the germinated pollen was firstly purified. And the vialbe sperm were released by a means of the grinding method. The viable sperm was purified with sugar density gradient centrifugation. The testing result shows that the isolated sperm cells might survive for 4h at room temperature.